Venezuelan (VEEV), eastern (EEEV), and western (WEEV) equine encephalitis viruses are arthropod-borne alphaviruses listed as CDC category B agents. There are currently no licensed human vaccines for the encephalitic alphaviruses, although conventional live-attenuated and formalin-inactivated vaccines for VEEV, EEEV, and WEEV have been developed and are currently utilized under IND status these vaccines have high reactogenicity and are poorly immunogenic. Ichor Medical Systems has demonstrated that a trivalent DNA vaccine candidate consisting of three DNA plasmids separately encoding the E1 and E2 glycoproteins of VEEV, EEEV and WEEV induce virus-neutralizing antibody responses consistent with protective immunity. Here we propose to further the development of this promising vaccine candidate by conducting IND-enabling safety/toxicology studies;this work encompasses both standard vaccine acute/chronic evaluation as well as cardiac monitoring to expand the safety database related to the clinical application of electroporation. In addition, preclinical immunogenicity experiments in nonhuman primates will be performed to facilitate the regulatory path toward approval of this product. PUBLIC HEALTH RELEVANCE: This SBIR proposes to conduct preclinical immunogenicity and safety/toxicology studies on a trivalent DNA vaccine against Venezuelan (VEEV), Eastern (EEEV), and Western (WEEV) equine encephalitis viruses. The work conducted in this project will enable clinical testing of a novel vaccine for an unmet medical need and further the development of electroporation as a broad vaccine delivery platform.